LOCUS       sequence-199470-        2474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of mOrange from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2474)
  AUTHORS   Stark JC, Huang A, Nguyen PQ, Dubner RS, Hsu KJ, Ferrante TC, 
            Anderson M, Kanapskyte A, Mucha Q, Packett JS, Patel P, Patel R, Qaq
            D, Zondor T, Burke J, Martinez T, Miller-Berry A, Puppala A, 
            Reichert K, Schmid M, Brand L, Hill LR, Chellaswamy JF, Faheem N, 
            Fetherling S, Gong E, Gonzalzles EM, Granito T, Koritsaris J, Nguyen
            B, Ottman S, Palffy C, Patel A, Skweres S, Slaton A, Woods T, 
            Donghia N, Pardee K, Collins JJ, Jewett MC
  TITLE     BioBits Bright: A fluorescent synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5107. doi: 10.1126/sciadv.aat5107. 
            eCollection 2018 Aug.
  PUBMED    30083609
REFERENCE   2  (bases 1 to 2474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5107. doi"; pages: " 
            10.1126/sciadv.aat5107. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1-mOrange
FEATURES             Location/Qualifiers
     source          1..2474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(220..239)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             660..682
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(713..732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(714..732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(760..1348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(840..859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1459..1478)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1470..2285
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1543..1562)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2303..2350)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2346..2364
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2416..2474,1..652))
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
ORIGIN
        1 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ccgacgatgt
       61 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcggaggtgt
      121 agtggccgcc gtccttcagc ttcagcctca tcttgatctc gcccttcagg gcgccgtcct
      181 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
      241 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
      301 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
      361 acttgaagcc ctcggggaag gacagcttga agtagtcggg gatgtcggcg gggtgcttca
      421 cgtaggcctt ggagccgtag gtgaactgag gggacaggat gtcccaggcg aagggcaggg
      481 ggccaccctt ggtcaccttc agcttagcgg tctgaaagcc ctcgtagggg cggccctcgc
      541 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgcgc accttgaagc
      601 gcatgaactc cttgatgatg gccatgttat tctcctcgcc cttgctcacc atatgtatat
      661 ctccttctta aagttaaaca aaattatttc tagagggaaa ccgttgtggt ctccctatag
      721 tgagtcgtat taatttcgcg ggatcgcgtt gctggcgttt ttccataggc tccgcccccc
      781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atctcacgtt gtgtctcaaa
     1381 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1441 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     1501 cgaagccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1561 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     1621 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1681 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1741 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     1801 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1861 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     1921 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     1981 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     2041 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2101 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2161 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2221 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2281 tctaatcaga attggctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     2341 gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt
     2401 tagcagccgg tcgacttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc
     2461 ggcgcgttcg tact
//
